Effect based monitoring tools Conclusions from Swedish workshop (January 2011) Ann-Sofie Wernersson (SE) Mario Carere (IT)

Effect based monitoring tools and evaluation criteria Jan 2011 Göteborg Sweden Aim: identify monitoring tools that can be used in Swedish limnic and marine surface water environments for the assessment of effects from hazardous substances. Organized by the County Administrative Board in Västra Götaland/Skagerrak and Kategat River Basin District, with financial contribution from the Swedish EPA as part of the project ”Coordinated system to assess the effects from hazardous substances in the aquatic environment” ( )

Outline 70 participants (national and international expertise) Upon registration, tools were “nominated” for evaluation 1st day: –15 flash presentations –presentation of similar French workshop on biomarkers (Ineris) –Four discussion groups: in vitro bioassays, biomarkers, in vivo bioassays and “higher organisational level tools”. Evaluation of selected tools according to questionnaire 2nd day: –related topics (TRIAD approach, mixture effects assessments, BLM) –Regulatory needs (WFD)

Flash presentations of tools –Biomarkers: fish biomarkers, reproduction effects on amphipods, diatom shell malformations, lysosomal stability in blue mussels, imposex, Chironomid mouth deformation –In vitro bioassays, incl Toxicity Profiling, Effects Directed Analysis –In vivo bioassays: Ceramium, Nitocra spinipes, FET (Fish Embryo Toxicity) –PICT (Pollution Induced Community Tolerance), SWIFT, SPEAR (SPEcies At Risk)

Tool specific discussions 4 discussion groups: –”in vitro”, ”single species in vivo”, ”biomarkers”, ”community level tools” Questionnaire (xls file), including the following questions: –Endpoint –Test duration –Suitable matrices (surface water, sediment etc) –Suitable in limnic, marine, brackish environments? –Species/cell type –References to studies that used the assay for monitoring purposes –Evaluation criteria available to evaluate results in absolute terms? –Possible to observe or predict effects on population and/or ecological level from test results? –Type of substances/modes of action the assay responds to –Highly specific or general? –Other factors that the assay might also respond to (e.g. oxygen, temperature etc) –Necessary to perform analysis during a certain season/avoid certain seasons? –Sensitivity of the assay? –Available commercial performers? –Known international regulatory requirement (e.g.dredged sedíment evaluation)? –Approximate costs –Complexity of analysis? –Additional research/validations needed before use within regular monitoring in Sweden?

Nominated tools acetylcholinesterase in vitro ALAD Ames umuC Micronucleus Comet Benthic diatoms benthic flux measurements Chronic sublethal toxicity tests on sediment dwelling organisms Daphnia magna diatom shell malformations dioxin-like effects, EROD FET-test (fish embryo toxicity) fish embryos, fish reproduction, fish behaviour, fish physiology Fish gill biomarker Fish Sexual Development Test Genetic adaptation and pollution tolerant ecotypes genotoxicity, health indices in mussels Heart rate as a sublethal indicator of stress in mussels Integrated (fish) monitoring LDH Lipid peroxidation Lysosome stability metallothionein microarrays, physiological aging and reproduction (fecundity, egg quality) microcosms Microtox, molecular tools, metagenomics Mouth deformation chironomids MTT Neutral red Oxidative damage physiological aging PICT reproduction (fecundity, egg quality) Reproductive success in fish SPEAR SWIFT Periphyton test The use of sticklebacks as a tool in biomonitoring TTR-binding assay and TH-responsive reporter gene assays (TR-CALUX?) V fisherii screening test

Evaluated tools Biomarkers: –Oxidative damage: protein carbonylation –Oxidative damage: lipid peroxidation –Oxidative damage: DNA oxidation –Oxidative damage: molecular antioxidants (i.e. GSH) –Lysosome stability –Scope for growth –Reproductive success in fish –EROD –Metallothionein –induction of vitellogenin in male fish –Diatoms shell malformations (biomarker?) Higher organisation levels –PICT –Swift –SPEAR in vitrobioassays: –AR CALUX –DR CALUX –ER CALUX –ERa CALUX –GR CALUX –PAH CALUX –PR CALUX –TRb CALUX –Acetylcholinesterase inhibition assay –Carboxylesterase inhibition assay –Ames fluctuation assay –umuC –TTR-binding In vivobioassays: –Ceramium tenuicorue –Fish embryo toxicity –Daphnia magna –Bacterial luminescence - Vibrio fischeri –Fish sexual development test –Nitocra spinipes

General conclusions: there are several effect based tools available for both marine and limnic Swedish environments for the study of several trophic levels (fish- invertebrates-plants) For different levels of organization (suborganism, individual, population, community) For several types of endpoints

Some tools need further development or validation studies. Development and validation needs especially for higher organisational level tools (PICT, SPEAR) Biomarkers: national baseline values and assessment criteria needed, especially for limnic environments Little national experience from using bioassays within monitoring except for investigative studies such as WEA (Whole Effluent Assessment). In particular little experience from in vitro bioassays within aquatic monitoring –Need to create baseline data.

Several potential uses of effect based monitoring tools also within the WFD Some disagreement about whether tools other than e.g. SPEAR and PICT truly measure effects on populations-communities –Would limit the use as biological quality elements within the WFD context. But agreed that several tools could be used to predict negative effects on population level –Would be analogous to the use of physicochemical quality elements (river basin specific pollutants) within ecological status classification

Documentation to be found Link to workshop agenda and presentations: dialog/seminarier-och-konferenser/Pages/default.aspx dialog/seminarier-och-konferenser/Pages/default.aspx French similar workshop: ineris/756http:// ineris/756 Upcoming report: ”Coordinated system to assess the effects from hazardous substances in the Swedish aquatic environment - review and recommendations” (prel title) –Will include annex on effect based tools, with aspects covered in questionnaires, but complemented with information from literature searches (expected in September 2011)